CN102993178A - Isoquinoline compound with Rho kinase inhibition activity and preparation method and application thereof - Google Patents
Isoquinoline compound with Rho kinase inhibition activity and preparation method and application thereof Download PDFInfo
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Abstract
The invention discloses an isoquinoline compound with Rho kinase inhibition activity and a preparation method and application thereof. The structure of the compound is shown by formula (I) or (II). The compound disclosed by the invention has good biological activity, strong effect on a target spot, strong cell regeneration and cynapse forming ability, high survival rate in a cell activity test and low death rate in a cell apoptosis test, and realizes lower toxicity of nitric oxide according to a test.
Description
Technical field
The invention belongs to biomedicine field, relate to isoquinoline compound, Preparation method and use with Rho kinase inhibiting activity.
Background technology
Cardiovascular and cerebrovascular diseases has become the first human killer, and people's life security and quality of life in serious threat.Research finds that the Rho kinases of activation can cause vasoconstriction, increased blood pressure, and amount of blood supply reduces, and the Rho kinases is one of important target spot for the treatment of cardiovascular and cerebrovascular diseases.Suppress the effective alleviate myocardial ischemia/reperfusion injury of the kinase whose intensity of activation of Rho, hypertension, cerebral apoplexy.In addition, evidence suggests that suppressing the activation in vivo of Rho kinases also has the growth of the nerve synapse of promotion, Spinal injury can be treated and alleviate to the effect of the neurological functional recovery after promoting to damage, alzheimer's disease, neural inflammation spinal cord takes off the nervus centralis diseases such as sheath.In addition, the infiltration of tumour and migration also depend on the kinase whose activation of Rho, suppress the transfer that the kinase whose activation of Rho can also suppress tumour.In sum, the kinase whose activation of inhibition Rho of efficient selective is very important.
The compound that can suppress the Rho kinase activator is referred to as the Rho kinase inhibitor.Rho kinases (Rho associatedcoil-coiled forming protein kinase, ROCK) has two hypotypes, i.e. ROCK1 and ROCK2, and they all are serine/threonine kinases.ROCK1 and ROCK2 are distributed widely in the most organs of body, and ROCK2 mainly is present in the brain, and that ROCK1 expresses in the organ of Non nervous system is higher, such as heart, and lung, skeletal muscle etc.The Rho kinases shrinks the division of cell, adheres to, and migration, the activities such as secretion have important regulating effect, such as smooth muscle contraction, Cytoskeleton, the growth of cell migration and proliferation and neuronic formation and aixs cylinder etc.
Even to this day, the Rho kinase inhibitor that is seen in report mainly contains iloquinoline derivative, indoles, acid amides and ureas and 4-aminopyridine class.The HA-1077(that belongs to iloquinoline derivative has another name called fasudil) as a kind of novel efficient vasodilator, can effectively alleviate cerebral vasospasm, improve Subarachnoid Hemorrhage (SAH) patient's prognosis.It enters clinically in the nineteen ninety-five official approval in Japan, and control chronic ischemic cerebral vasospasm is the Rho kinase inhibitor of so far unique listing, has broad application prospects.
But in the use procedure of fasudil, metabolism is fast in vivo to it is found that it, and is difficult for seeing through hemato encephalic barrier.For this reason the present invention design, synthetic and give birth to surveyed a series of isoquinoline compounds after screening obtained two stronger to the Rho kinase inhibiting activity, toxic side effect is lower, has certain patent medicine prospect.
Summary of the invention
Primary and foremost purpose of the present invention is that metabolism is fast in vivo in order to overcome existing fasudil, is difficult for seeing through the deficiency of hemato encephalic barrier, provides to have Rho kinase inhibiting activity height and the low isoquinoline compound of toxic side effect.
Second purpose of the present invention provides the preparation method of the isoquinoline compound with Rho kinase inhibiting activity.
The 3rd purpose of the present invention provides the purposes of the isoquinoline compound with Rho kinase inhibiting activity.
Technical scheme of the present invention is summarized as follows:
Isoquinoline compound with Rho kinase inhibiting activity represents with formula (I) or (II):
The preparation method of compound (I) comprises the steps:
Under ice bath, with L-dried meat ammonia alcohol and the reaction of 5-chlorosulfonyl isoquinoline 99.9, generate the isoquinoline compound (I) with Rho kinase inhibiting activity.
The preparation method of compound (II) comprises the steps:
(1) under ice bath, with L-dried meat ammonia alcohol and the reaction of 5-chlorosulfonyl isoquinoline 99.9, generates the isoquinoline compound (I) with Rho kinase inhibiting activity;
(2) compound (I) is dissolved in the thionyl chloride, in 60 ℃ of reactions 7 hours, obtains having the isoquinoline compound (II) of Rho kinase inhibiting activity.
Compound (I) or compound (II) purposes in preparation inhibition Rho kinase activity medicine.
The isoquinoline compound activity with Rho kinase inhibiting activity that the present invention is prepared is good, effect to target spot is stronger, it is strong that cell regeneration and cynapse form ability, high to surviving rate in the cytoactive, mortality ratio is low in the apoptosis test, and the nitrogen protoxide toxotest is lower, has very good patent medicine prospect.
Description of drawings
Fig. 1 is the mensuration of Rho kinase inhibitor activity.
Fig. 2 is the test (BV2 cell) that cell regeneration and cynapse form ability.
Fig. 3 is that cynapse forms aptitude tests (BV2 cell).
Fig. 4 is that cell regeneration and cynapse form aptitude tests (primary neuron cell).
Fig. 5 is the cytoactive test.
Fig. 6 is the apoptosis test.
Fig. 7 is the nitric oxide concentration test.
Embodiment
This experimental design, synthetic and detected 9 isoquinoline compounds, screening obtained two stronger to the Rho kinase inhibiting activity, toxic side effect is lower, has certain patent medicine prospect.
The below lists the building-up process of these nine compounds one by one:
Synthesizing of embodiment 1 5-(4-benzyl-Isosorbide-5-Nitrae-Diazesuberane-1-alkylsulfonyl) isoquinoline 99.9 (No. 1 compound)
Fasudil hydrochloride (1.00g, 2.74mmol) is dissolved in the 10mL water, adds the 5mL aqueous solution that is dissolved with salt of wormwood (0.68g, 4.93mmol), after mixing, obtain pale yellow solution, use again CH
2Cl
2Extraction (50mL * 2) merges organic phase, and anhydrous magnesium sulfate drying filters, and behind the concentrating under reduced pressure, and triethylamine (0.33g, 3.30mmol) is dissolved in 50mLCH jointly
2Cl
2In, then to wherein adding benzyl chlorine (0.42g, 3.29mmol), stir reaction 30h under the room temperature.Pressure reducing and steaming CH
2Cl
2, column chromatography [the V(sherwood oil): the V(ethyl acetate)=1:3] separate and to obtain 0.40g yellow oil, yield 38.1%.
Gained compound chemistry shift value is as follows:
1H-NMR（500MHz，CDCl
3）δ：1.80～1.85（m，2H），2.65～2.68（m，4H），3.47～3.51（m，4H），3.59（s，2H），7.20～7.29（m，5H），7.65～7.68（m，1H），8.16（d，J＝9.0Hz，1H），8.33（d，J＝7.5Hz，1H），8.45（d，J＝6.0Hz，1H），8.66（d，J＝6.0Hz，1H），9.33（s，1H）。
Synthesizing of embodiment 2 5-(4-methoxy carbonyl ethyl-Isosorbide-5-Nitrae-Diazesuberane-1-alkylsulfonyl) isoquinoline 99.9 (No. 2 compounds)
Fasudil hydrochloride (2.00g, 5.49mmol) is dissolved in the 10mL water, adds the 5mL aqueous solution that is dissolved with salt of wormwood (1.36g, 9.86mmol), after mixing, obtain pale yellow solution, use again CH
2Cl
2Extraction (50mL * 2) merges organic phase, and anhydrous magnesium sulfate drying filters, and behind the concentrating under reduced pressure, is dissolved in 50mL CH
2Cl
2In, then to wherein adding methyl acrylate (1.42g, 16.47mmol), stir reaction 30h under the room temperature.Pressure reducing and steaming CH
2Cl
2, column chromatography (pure ethyl acetate) separation obtains 1.65g light yellow oil, yield 79.6%.
Gained compound chemistry shift value is as follows:
1H-NMR(500MHz, CDCl
3) δ: 1.78～1.82(m, 2H), 2.65～2.68(m, 4H), 3.32, (s, 2H), 3.47～3.51(m, 4H), 3.68(s, 3H), 7.20～7.29(m, 5H), 7.65～7.68(m, 1H), 8.16(d, J=9.0Hz, 1H), 8.33(d, J=7.5Hz, 1H), 8.45(d, J=6.0Hz, 1H), 8.66(d, J=6.0Hz, 1H), 9.33(s, 1H).
Synthesizing of embodiment 3 5-(o-chlorobenzyl-Isosorbide-5-Nitrae-Diazesuberane-1-alkylsulfonyl) isoquinoline 99.9 (No. 3 compounds)
Fasudil hydrochloride (2.00g, 5.49mmol) is dissolved in the 10mL water, adds the 5mL aqueous solution that is dissolved with salt of wormwood (1.36g, 9.86mmol), after mixing, obtain pale yellow solution, use again CH
2Cl
2Extraction (50mL * 2) merges organic phase, and anhydrous magnesium sulfate drying filters, and behind the concentrating under reduced pressure, and triethylamine (0.66g, 6.59mmol) is dissolved in 50mLCH jointly
2Cl
2In, then to wherein adding o-chlorobenzyl chlorine (2.66g, 16.47mmol), stir reaction 30h under the room temperature.Pressure reducing and steaming CH
2Cl
2, column chromatography (pure ethyl acetate) separation obtains 1.94g light yellow oil, yield 85.1%.
Gained compound chemistry shift value is as follows:
1H-NMR（500MHz，CDCl
3）δ：1.81～1.85（m，2H），2.66～2.69（m，4H），3.50～3.53（m，4H），3.59（s，2H），7.23～7.27（m，4H），7.65～7.68（m，1H），8.18（d，J＝9.0Hz，1H），8.36（d，J＝7.5Hz，1H），8.45（d，J=6.0Hz，1H），8.66（d，J=6.0Hz，1H），9.36（s，1H）。
Fasudil hydrochloride (1.00g, 2.74mmol) is dissolved in the 10mL water, adds the 5mL aqueous solution that is dissolved with salt of wormwood (0.68g, 4.93mmol), after mixing, obtain pale yellow solution, use again CH
2Cl
2Extraction (50mL * 2) merges organic phase, and anhydrous magnesium sulfate drying filters, and behind the concentrating under reduced pressure, and triethylamine (0.33g, 3.29mmol) is dissolved in 50mLCH jointly
2Cl
2In, then to wherein adding p-chlorobenzyl chlorine (1.33g, 8.23mmol), stir reaction 30h under the room temperature.Pressure reducing and steaming CH
2Cl
2, column chromatography (pure ethyl acetate) separation obtains 0.70g yellow oil, yield 61.4%.
Gained compound chemistry shift value is as follows:
1H-NMR（500MHz，CDCl
3）δ：1.81～1.86（m，2H），2.65～2.68（m，4H），3.47～3.52（m，4H），3.56（s，2H），7.17～7.18（m，2H），7.24～7.27（m，2H）7.67～7.70（m，1H），8.19（d，J＝8.0Hz，1H），8.33～8.35（m，1H），8.45（d，J＝7.5Hz，1H），8.68（d，J＝6.0Hz，1H），9.35（s，1H）。
Get the hydrochloride (1.00g, 3.79mmol) of isoquinoline 99.9-5-SULPHURYL CHLORIDE, be dissolved in the 50mL water, stir the lower saturated sodium bicarbonate solution that drips, regulate pH=5 ~ 6, use CH
2Cl
2Extraction (50mL * 2) merges organic phase, and anhydrous sodium sulfate drying is concentrated into 50mL, then to wherein adding the CH that contains 2,2,6,6-tetramethyl piperidine amine (1.17g, 7.576mmol)
2Cl
2Solution 20mL, stirring reaction 4h under room temperature, CH is removed in underpressure distillation
2Cl
2, column chromatography [V(methyl alcohol): the V(ethyl acetate)=1:1] separate, get faint yellow oily thing 0.23g, yield 17.4%.
Gained compound chemistry shift value is as follows:
1H-NMR（500MHz，CDCl
3）δ：0.97（s，6H），1.03（s，6H），1.24～1.28（m，2H），1.54～1.58（m，2H），3.47～3.62（m，1H），5.10（s，1H），7.70～7.73（m，1H），8.22（d，J＝10.0Hz，1H），8.44（d，J＝6.0Hz，1H），8.48～8.50（m，1H），8.68（d，J＝6.0Hz，1H），9.38（s，1H）。
Synthesizing of embodiment 6 5-(4-hydroxyethyl-Isosorbide-5-Nitrae-Diazesuberane-1-alkylsulfonyl) isoquinoline 99.9 (No. 6 compounds)
Fasudil hydrochloride (1.00g, 2.74mmol) is dissolved in the 10mL water, adds the 5mL aqueous solution that is dissolved with salt of wormwood (0.68g, 4.93mmol), after mixing, obtain pale yellow solution, use again CH
2Cl
2Extraction (50mL * 2) merges organic phase, and anhydrous magnesium sulfate drying filters, and behind the concentrating under reduced pressure, and triethylamine (0.33g, 3.29mmol) is dissolved in 50mLCH jointly
2Cl
2In, then to wherein adding bromoethanol (0.90g, 8.23mmol), stir reaction 30h under the room temperature.Pressure reducing and steaming CH
2Cl
2, column chromatography (pure ethyl acetate) separation obtains 0.57g yellow oil, yield 65.1%.
1H-NMR（500MHz，CDCl
3）δ：1.81～1.86（m，2H），2.61(t，J
1=6.0HZ，J
2=6.0Hz，2H)，2.65～2.68（m，4H），3.47～3.51（m，4H），3.59（m，2H），7.65～7.68（m，1H），8.17（d，J＝9.0Hz，1H），8.34（d，J＝7.5Hz，1H），8.45（d，J＝6.0Hz，1H），8.66（d，J＝6.0Hz，1H），9.33（s，1H）。
Synthesizing of embodiment 7 5-(1-benzylamino-Isosorbide-5-Nitrae-Diazesuberane-1-alkylsulfonyl) isoquinoline 99.9 (No. 7 compounds)
Get the fasudil oxynitride (2.00g, 4.53mmol) of Cbz protection, be dissolved in 30mLCH
2Cl
2In, under ice bath, drip phosphorus oxychloride 5mL, back flow reaction 7h contains the 25%NaOH solution of frozen water with the reaction solution impouring, vigorous stirring, and with 5% NaOH solution regulator solution to pH=10, tell CH
2Cl
2Phase, water CH
2Cl
2Extraction (50mL * 2) merges organic phase, and drying is filtered, and obtains the deep yellow viscous fluid after the underpressure distillation, and column chromatography [the V(sherwood oil): the V(ethyl acetate)=1:1] separate and obtain white solid 1.47g, yield 70.7%.
Above-mentioned white solid (1.00g, 2.17mmol) is dissolved in the 15mL benzylamine, then in 70 ℃ of reaction 3h, add large water gaging, then 1MHCl transfers pH=7, ethyl acetate extraction, and drying is filtered, the pressure reducing and steaming ethyl acetate obtains light green oily matter 1.00g, yield 87.0%.
Get above-mentioned light green oily matter (1.00g, 1.89mmol) and be dissolved in the 50mL ethyl acetate, then add the 0.20g palladium-carbon catalyst, stirring at room reaction 24h, steam ethyl acetate, column chromatography [the V(sherwood oil): the V(ethyl acetate)=1:3] light green oily matter 0.41g, yield 56.9%.
Gained compound chemistry shift value is as follows:
1H-NMR（500MHz，CDCl
3）δ：1.99～2.04（m，2H），3.25～3.28（m，4H），3.43（t，J
1=6.0Hz，J
2=6.0Hz，2H），3.62（t，J
1=6.0Hz，J
2=6.0Hz，2H），4.75（s，2H），7.26～7.32（m，5H）,7.47（d，J＝7.5Hz，1H），7.59（d，J＝8.5Hz，1H），7.72～7.75（m，1H），8.29～8.30（m，1H），8.50（d，J＝6.0Hz，1H）。
(1) vitriol (10.00g, 32.60mmol) with 5-sulfonic acid isoquinoline 99.9 is dissolved in the sulfur oxychloride (75.00g, 630mmol), drips simultaneously DMF (10mL), and mechanical stirring is in 60 ℃ of reaction 5h.Unnecessary sulfur oxychloride is removed in distillation, then adds CH
2Cl
2(200mL), separate out a large amount of crystal, filter filter cake CH
2Cl
2Washing, the vacuum Air drying gets 9.60g white powder 5-chlorosulfonyl isoquinoline hydrochloride, and yield is 90.5%.
(2) above-mentioned white powder (2.00g, 7.60mmol) is added in the frozen water (25mL), stirring lower is 5-6 with saturated sodium bicarbonate solution adjusting pH value, CH
2Cl
2Extraction (50mL * 2) merges organic phase, and anhydrous sodium sulfate drying filters, and filtrate is concentrated into 50mL (5-chlorosulfonyl isoquinoline 99.9) for subsequent use.
(3) L-dried meat ammonia alcohol (0.92g, 9.10mmol) is dissolved in CH
2Cl
2(50mL), under 0 ~ 5 ℃, to the solution that wherein drips step (2) acquisition, finish room temperature reaction 2h.Evaporate to dryness CH
2Cl
2, add residuum with ethyl acetate and water, tell organic phase, concentrated, get the 2.12g light yellow oil.Yield is 95.9%, HPLC purity〉99%.
Gained compound chemistry shift value is as follows:
1HNMR(400MHz，CDCl
3)δ：9.42(s，1H)，8.74(s，2H)，8.49(d，J＝7.3Hz，1H)，8.29(d，J＝8.1Hz，1H)，7.78(t，J＝7.8Hz，1H)，3.97~3.82(m，1H)，3.71(q，J
1=11.4，J
2=4.9Hz，2H)，3.56~3.32(m，2H)，1.99~1.70(m，3H)，1.66~1.51(m，1H)。
Its structural formula is:
Compound (I)
Embodiment 9 5-'s (2-chloromethyl-tetramethyleneimine-1-alkylsulfonyl)-isoquinoline 99.9 (No. 9 compounds, i.e. compound (II)) is synthetic
Compound (I) (1.60g, 5.50mmol) directly is dissolved in the 25mL thionyl chloride, and in 60 ℃ of reactions 7 hours, then the excessive thionyl chloride of pressure reducing and steaming added 50mL water, saturated NaHCO
3Solution is regulated pH=6.0, and dichloromethane extraction (50mL * 3) merges organic phase, and drying is filtered, and the pressure reducing and steaming solvent gets faint yellow oily thing 1.20g after the column chromatography for separation, and yield is 70.6%, HPLC〉98%.
Gained compound chemistry shift value is as follows:
1H?NMR(400MHz，CDCl
3)δ：9.45(s，1H)，8.93(d，J=5.8Hz，1H)，8.72(s，1H)，8.61(d，J=7.3Hz，2H)，7.92(t，J=7.4Hz，1H)，4.14(s，1H)，3.78(d，J＝11.0Hz，1H)，3.66~3.50(m，H)，3.40(d，J=16.5Hz，2H)，1.99~1.70(m，3H)，1.66~1.51(m，1H)。
Its structural formula is:
Compound (II)
The HPLC condition:
Be weighting agent with 18 silane glue: the 0.20g SODIUM PHOSPHATE, MONOBASIC adds 500mL distilled water, splashes into 2 phosphoric acid, control pH=3.0) be buffering salt; Methyl alcohol: buffering salt=7:3 is moving phase, and the detection wavelength is 274nm, and flow velocity is 1.0mL/min, and theoretical plate number should be calculated by Fasudil derivative and be no more than 4000.
(1) product of embodiment 1-9 preparation is an amount of respectively, with the methyl alcohol dilution, is mixed with the 0.2mg/mL sample solution, gets 20ul and injects sample introduction, detects under above-mentioned HPLC condition.
(2) analytical results: shown in the product liquid phase spectrogram of embodiment 1-9 preparation: each product content is all greater than 99%.
The product of embodiment 1-9 preparation is used for the Rho kinase inhibiting activity, the test of apoptosis and cynapse regeneration, the structure of compound is as follows.The order of embodiment is corresponding one by one with the order of compound.
Embodiment 10 biological activity tests
10.1Rho the test of kinase inhibitor activity
Microglia BV2 is as the immunocyte in the central nervous system; both can neurone be shielded by pathogenic agent and the deleterious particle of engulfing in the cerebral tissue; again can be under the effect of pro-inflammatory cytokine; activate into microglia of reactivity; secrete inflammatory cytokines plays toxic action to neurone, is an important target spot of the neural inflammation for the treatment of and nerve degenerative diseases.By the BV2 microglia respectively with the product effect of fasudil, embodiment 1-9 preparation after 24 hours cell culture supernatant Rho kinase activity measure, its result is as shown in Figure 1.
The BV2 microglia sample that PBS processes is compared, and except 3, No. 7 compounds do not suppress beyond the activity the Rho kinases, other compounds can both show certain Rho kinase inhibiting activity, wherein with 6,8, and No. 9 compound optimums.
10.2 cell regeneration and cynapse form the test of ability
Fig. 2 be the BV2 microglia respectively with the product effect of fasudil, embodiment 1-9 preparation microscope document image after the fixing washing of cell after 48 hours.The BV2 spongiocyte contrast that PBS processes, fasudil, No. 8 compounds and No. 9 compounds have obvious cell regeneration and cynapse formation ability, and 1,2, No. 6 compound takes second place.Compare with fasudil, No. 8 compounds and No. 9 compounds have cell regeneration and the cynapse stronger than fasudil and form ability.
Fig. 3 be the BV2 microglia respectively with the product effect of fasudil, embodiment 1-9 preparation after cynapse formational situation under 24 hours cultivation conditions.Relatively PBS processes, and fasudil and the obvious cynapse of demonstration form ability.2 and 6 compounds take second place.
Fig. 4 is that former culture neurone is respectively with the product effect of fasudil, embodiment 1-9 preparation after 48 hours after the fixing washing of cell, through monoclonal anti neuronal antibodies dyeing microscope document image.Relatively PBS processes, and fasudil and 8, No. 9 identical and obvious cell regeneration of compound exhibits and cynapse form ability.2 and No. 6 compound takes second place, and all the other compounds are all relatively poor.
To sum up, 8 and No. 9 compounds have obvious cell regeneration and cynapse formation ability.2 and No. 6 compound takes second place.
10.3 cytoactive and apoptosis test
Active for test cell, adopt the BV2 microglia to measure by mtt assay, by the apoptosis of LDH test cell.
Mtt assay is mainly for detection of the method for cell survival and growth.The ultimate principle of its effect is as follows: the desaturase in the viable cell plastosome can reduce yellow bromination 3-(4,5-dimethylthiazole-2)-2, [3-(4 for 5-phenylbenzene four nitrogen, 5-dimethylthiazol-2yl)-2,5-diphenylterazolium bromide, MTT] be hepatic water-fast formazan, generation De formazan can be deposited in the viable cell, and dead cell is then without this function.With the dissolving of the formazan in the cell, under the wavelength of 490nm measure its light absorption value (OD value) with enzyme-linked immunosorbent assay instrument with dimethyl sulfoxide (DMSO), can indirectly reflect viable cell quantity.
Fig. 6 be the BV2 microglia respectively with the product effect of fasudil, embodiment 1-9 preparation after 48 hours, cell MTT measurement result.Relatively PBS processes, and can find out and only have preferably cytoactive of fasudil and 8 and No. 9 compound exhibits, and other compounds has the effect of obvious inhibition cytoactive.
LDH tests serum lactic dehydrogenase exactly, and the LDH test is exactly the concentration of measuring LDH.In the LDH antibody of purifying, add test-compound, through thoroughly developing the color with substrate tetramethyl benzidine (TMB) after the washing, TMB changes into blueness under the catalysis of serum lactic dehydrogenase, and under the effect of acid, finally change into yellow, under the wavelength of 450nm, measure light absorption value (OD value) with microplate reader, the LDH concentration positive correlation in the depth of color (size of OD value) and the sample.
Fig. 7 be the BV2 microglia respectively with the product of fasudil, embodiment 1-9 preparation cell culture supernatant LDH measurement result after 24 hours.With result's contrast that PBS processes, fasudil and 8 and No. 9 compound exhibits LDH release are lower, show that the necrocytosis number is lower, and 2 and No. 6 compounds take second place, and other compound then has stronger cell lethality.
The MTT cytoactive of 8 and No. 9 compounds does not reduce, and the apoptosis number of LDH is few, and namely for the toxicity of cell, 8 and No. 9 compound is less, and 2 and No. 6 compound takes second place.
10.4 nitric oxide concentration test
Nitrogen protoxide has participated in all too many levels such as genesis of ischemic cerebrovascular, and the NO of high density can the direct killing neurocyte.Figure 8 shows that BV2 microglia method respectively with the product effect of fasudil, embodiment 1-9 preparation after the result that measures of 24 hour cell culture supernatant nitric oxide concentrations.Relatively PBS processes, fasudil and 6,8, and No. 9 compound all has obvious inhibition; Compare fasudil, 6,8, No. 9 compound exhibits go out the effect of the inhibition NO stronger than fasudil.6,8, No. 9 compound exhibits goes out the neurocyte protection effect stronger than fasudil.
8 and No. 9 compound has the Rho kinase inhibiting activity stronger than fasudil, and has the Rho kinase inhibiting activity, and lower side effect, and 2 and No. 6 compounds take second place.
Claims (4)
1. the isoquinoline compound that has the Rho kinase inhibiting activity, its structure are suc as formula (I) or (II):
2. the preparation method of the compound of claim 1 (I), its feature comprises the steps:
Under ice bath, with L-dried meat ammonia alcohol and the reaction of 5-chlorosulfonyl isoquinoline 99.9, obtain having the isoquinoline compound (I) of Rho kinase inhibiting activity.
3. the preparation method of the compound of claim 1 (II), its feature comprises the steps:
(1) under ice bath, with L-dried meat ammonia alcohol and the reaction of 5-chlorosulfonyl isoquinoline 99.9, generates the isoquinoline compound (I) with Rho kinase inhibiting activity;
(2) compound (I) is dissolved in the thionyl chloride, in 60 ℃ of reactions 7 hours, obtains having the isoquinoline compound (II) of Rho kinase inhibiting activity.
4. compound described in the claim 1 suppresses the purposes of Rho kinases medicine in preparation.
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| WO2015165341A1 (en) * | 2014-04-28 | 2015-11-05 | 南京明德新药研发股份有限公司 | Isoquinolinesulfonyl derivative as rho kinase inhibitor |
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| CN101622243A (en) * | 2007-02-28 | 2010-01-06 | 旭化成制药株式会社 | Sulfonamide derivative |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| WO2015165341A1 (en) * | 2014-04-28 | 2015-11-05 | 南京明德新药研发股份有限公司 | Isoquinolinesulfonyl derivative as rho kinase inhibitor |
| JP2017513919A (en) * | 2014-04-28 | 2017-06-01 | メッドシャイン ディスカバリー インコーポレイテッド | Isoquinoline sulfone derivatives as RHO kinase inhibitors |
| US9856264B2 (en) | 2014-04-28 | 2018-01-02 | Medshine Discovery Inc. | Isoquinolinesulfonyl derivative as RHO kinase inhibitor |
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